134 



LABORATORY COURSE IN SERUM STUDY 



II 



GLANDERS COMPLEMENT FIXATION 



This is carried out in the same manner as the gonococcus test, 

 using the serum of a known glanders-infected horse as a positive con- 

 trol. 1 



Bring up to 2.0 c.c. with salt solution. 



Incubate one hour in the thermostat at 37 C. or if possible four 

 hours in the ice box at 10 C. Then add 0.5 c.c. sheep cells sensitized 

 with two units of amboceptor and incubate at 37 C. until all tubes 

 .excepting Tube 1 are. completely laked. The positive serum is from a 

 glandered horse known to be positive by previous tests. 



Materials needed for each student for gonococcus fixation : 



0.5 c.c. positive serum. 



0.5 c.c. normal serum. 



5.0 c.c. antigen (diluted). 



5.0 c.c. guinea pig serum (1-10). 



5.0 c.c. shesp cells 5 per cent suspension. 



20 units amboceptor. 

 20 half-inch test tubes 



Salt solution. 

 1 test tube rack. 

 lOone-c.c. pipettes (graduated). 



GLANDERS ANTIGEN 



1 The technique of preparing glanders antigen as practiced at the New York 

 Board of Health and supplied to us by the kindness of Miss M. P. Olmstead is as 

 follows : 



Stock transplants are kept on glycerine potato agar (for formula see Park 

 and Williams' Pathogenic Microorganisms, 1914, page 97) from 1 per cent to 

 2 per cent acid to phenolphthalein. For antigen transplants the same medium 

 is used as in the preparation of gonococcus antigen, except for the reaction, which 

 should be 1.6 per cent acid. The 24 hour growth on bottles is washed off with 

 sterile, neutral, distilled water and heated at 80 C. for 6 to 8 hours, then filtered. 

 With the exceptions mentioned, the technique of antigen preparation is the same 

 as in the case of gonococcus. 



